In:
Journal of AOAC INTERNATIONAL, Oxford University Press (OUP), Vol. 98, No. 1 ( 2015-01-01), p. 1-4
Abstract:
Development of agricultural biotechnology requires rapid and convenient methods for crop plant genotyping. Real-time PCR is sensitive and reliable, and has been a routine technique in plant research. However, its application is limited by the cumbersome DNAtemplate preparation procedures. We tested three PCRmaster mixes for direct amplification of crude seed DNA extracts without extensive purification. One mixhad higher resistance to plant-derived PCR inhibitors and was shown to be applicable to various important crop plants. Furthermore, this method is capable of detecting single-copy genes from 2 mg pieces of seeds repetitively. Meanwhile, melting curve analysis could detect amplicons directly without electrophoresis manipulations. Taken together, this direct real-time PCR method provides a rapid and convenient toolfor seed genotypic screening in crop plants.
Type of Medium:
Online Resource
ISSN:
1060-3271
,
1944-7922
DOI:
10.5740/jaoacint.14-104
Language:
English
Publisher:
Oxford University Press (OUP)
Publication Date:
2015
